ABSTRACT
ABSTRACT The aim of the present study was to analyze the chemical composition ofCurcuma longa,Pimenta dioica,Rosmarinus officinalis, andSyzygiumaromaticumessential oils (EOs) and their antifungal and anti-conidiogenic activity againstFusarium verticillioides. The chemical profile of the EOs was determined by GC/MS. The antifungal and anti-conidiogenic activities were evaluated by the agar dilution method. The tested concentrations were 1000ppm, 500ppm, 250ppm and 125ppm.S. aromaticumEO exhibited the highest antifungal effect, followed byP. dioicaand to a lesser extentC. longa. The major compounds of these EOs were eugenol (88.70% inS. aromaticumand 16.70% inP. dioica), methyl eugenol (53.09% inP. dioica), and α-turmerone (44.70%), β-turmerone (20.67%), and Ar-turmerone (17.27%) inC. longa.Rosmarinus officinalispoorly inhibited fungal growth; however, it was the only EO that inhibited conidial production, with its major components being 1,8-cineole (53.48%), α-pinene (15.65%), and (−)-camphor (9.57%). Our results showed that some compounds are capable of decreasing mycelial growth without affecting sporulation, and vice versa. However, not all the compounds of an EO are responsible for its bioactivity. In the present work, we were able to identify different major compounds or mixtures of major compounds that were responsible for antifungal and anti-conidiogenic effects. Further experiments combining these pure components are necessary in order to achieve a highly bioactive natural formulation against the phytopathogenic fungusF. verticillioides.
RESUMEN El objetivo del presente estudio fue analizar la composición química de los aceites esenciales (AE) deCurcuma longa,Pimenta dioica,RosmarinusofficinalisySyzygiumaromaticum, y su actividad antifúngica y anti-esporuladora contraFusarium verticillioides. La composición de los AE se analizó por CG-EM. La actividad antifúngica y anti-esporuladora se evaluó a través del método de dilución en agar usando las siguientes concentraciones: 1.000, 500, 250 y 125ppm. El AE deS. aromaticumdemostró el mayor efecto antifúngico, seguido del deP. dioicay, en menor medida, delC. longa. Los compuestos principales de estos AE fueron eugenol (88,70% enS. aromaticumy 16,70% enP. dioica), metileugenol (53,09% enP. dioica) y α-turmerona (44,70%), β-turmerona (20,67%) y Ar-turmerona (17,27%) enC. longa. El AE deR. officinalisfue el que menor efecto inhibitorio presentó sobre el crecimiento fúngico, pero fue el único que inhibió la producción de conidias; sus principales componentes fueron 1,8-cineol (53,48%), α-pineno (15,65%) y (−)-alcanfor (9,57%). Nuestros resultados demostraron que algunos compuestos son capaces de disminuir el crecimiento del micelio deF. verticillioidessin afectar la esporulación, y vice versa. Sin embargo, no todos los compuestos de un AE son responsables de su bioactividad. En el presente trabajo, pudimos identificar diferentes compuestos o mezclas de compuestos que fueron responsables de los efectos antifúngicos y anti-esporuladores. Se necesitan nuevos experimentos que evalúen la combinación de estos compuestos puros para lograr una formulación bioactiva y de origen natural para el control deF. verticillioides.
Subject(s)
Oils, Volatile , Fusarium , Oils, Volatile/pharmacology , Microbial Sensitivity Tests , Zea mays , Antifungal Agents/pharmacologyABSTRACT
Abstract Currently, the valorization of agroindustrial waste is of great interest. Moringa oleifera is a multipurpose tree whose softwood residues could be used as raw material for low-cost cellulase production. The aim of this study was to isolate, identify, and characterize microorganisms with cellulolytic activity in different carbon sources. We isolated and puri-fied 42 microorganisms from M. oleifera biomass. Fungi presenting the largest hydrolytic halos in carboxymethylcellulose as a substrate were molecularly identified as Penicillium funiculosum (FG1), Fusarium verticillioides (FG3) and Cladosporium cladosporioides (FC2). The ability of these fungal strains to break down cellulose was assessed in a submerged fermentation using either amorphous CMC or crystalline form (Avicel). P. funiculosum and C. cladosporioides displayed similar endoglucanase (606 U/l) and exoglucanase (205 U/l) activities in the Avicel-containing medium, whereas F. verticillioides showed the highest level of p-glucosidase activity (664 U/l) in the carboxymethylcellulose medium. In addition, the effect of three culture media (A, B, and C) on cellulase production was evaluated in P. funiculosum using moringa straw as a carbon source. The results showed a volumetric productivity improvement of cellulases that was 2.77-, 8.26-, and 2.30-fold higher for endoglucanase, exoglucanase and p-glucosidase, respectively when medium C containing moringa straw was used as a carbon source. The enzymatic extracts produced by these fungi have biotechnological potential especially for second-generation bioethanol production (2G) from moringa straw. This is the first report on the use of M. oleifera biomass to induce the production of various cellulases in P. funiculosum. © 2019 Asociación Argentina de Microbiología. Published by Elsevier Espana, S.L.U. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/).
Resumen Actualmente, la valorización de los residuos agroindustriales es de gran interés. En este trabajo se emplearon residuos de madera blanda de Moringa oleifera para la producción de celulasas de bajo costo. El objetivo fue aislar, identificar y caracterizar microorganismos con actividad celulolítica en diferentes fuentes de carbono. A partir de la biomasa de M. oleifera, se aislaron e identificaron 42 microorganismos productores de celulasas. Los hongos que presentaron los mayores halos de hidrólisis en carboximetilcelulosa como sustrato fueron identificados molecularmente como Penicillium funiculosum (FG1), Fusarium verticillioides (FG3) y Cladosporium cladosporioides (FC2). Mediante fermentación sumergida, se evaluó la capacidad de estas cepas en la producción de celulasas utilizando celulosa cristalina (Avicel) y amorfa (CMC) como fuentes de carbono. P. funiculosum y C. cladosporioides presentaron las mayores actividades de endoglucanasa (606 U/l) y exoglucanasa (205 U/l) en medio Avicel, mientras que F. verticillioides mostró la mayor actividad de p-glucosidasa (664 U/l) en medio CMC. Además, se evaluó el efecto de tres medios de cultivo (A, B y C) sobre la producción de celulasas en P. funiculosum empleando residuos de moringa como fuente de carbono. Los resultados mostraron que en el medio C, la productividad volumétrica de celulasas se incrementó en 2,77; 8,26 y 2,30 veces para las actividades de endoglucanasa, exoglucanasa y p-glucosidasa, respectivamente. Los extractos enzimáticos producidos tienen gran potencial para su utilización biotecnológica, especialmente en la sacarificación de residuos de moringa y la producción de bioetanol de segunda generación. Este es el primer estudio del uso de la biomasa de M. oleifera para inducir la producción de diversas celulasas en P. funiculosum.
Subject(s)
Cellulase/physiology , Cellulose/metabolism , Cladosporium/enzymology , Moringa oleifera/enzymology , Talaromyces/enzymology , Fusarium/enzymologyABSTRACT
Endophytic fungi are a promising source for discovery of compounds with biotechnological potential. The aim of this study was to select and identify endophytic fungi from Coffea arabica that produce volatile organic compounds (VOCs), evaluate the effect of the VOCs produced by endophytic fungi on the growth of Rhizoctonia solani, Fusarium oxysporum, Phoma sp., Botrytis cinerea, Fusarium solani, Fusarium verticillioides, Cercospora coffeicola and Pestalotia longisetula, and select endophytic fungi with potential for biological control of Aspergillus ochraceus inoculated in coffee beans and F. verticillioides inoculated in corn seeds. An isolate of Muscodor albus was used as selection tool for VOC producing fungi. Among the 400 endophytic fungi isolates, 11 were able to grow in the presence of VOCs produced by M. albus. These fungi were identified as Muscodor spp. (9) and Simplicillium sp. according to searches in UNITE database using DNA sequences of internal transcribed spacer (ITS). The VOC's produced by endophytic fungi inhibited the growth the phytopathogenic fungi with different efficacies, compared to the control. The VOCs produced by Muscodor coffeanum (COAD 1842) showed fungicidal effect against A. ochraceus on coffee beans. Six endophytic fungi completely inhibited growth of F. verticillioides inoculated in corn seeds. This study demonstrates that the volatile-compound producing endophytic fungi, isolated from Coffea arabica, are promising sources of bioactive compounds.
Fungos endofíticos são uma fonte promissora para a descoberta de compostos com potencial biotecnológico. O objetivo deste estudo foi selecionar e identificar fungos endofíticos de Coffea arabica que produzem compostos orgânicos voláteis (COVs), avaliar o efeito dos compostos orgânicos voláteis produzido por fungos endofíticos sobre o crescimento de Rhizoctonia solani, Fusarium oxysporum, Phoma sp., Botrytis cinerea, Fusarium solani, Fusarium verticillioides, Cercospora coffeicola e Pestalotia longisetula e selecionar fungos endofíticos com potencial para controle biológico de Aspergillus ochraceus inoculado em grãos de café e F. verticillioides inoculado em sementes de milho. Um isolado de Muscodor albus foi utilizado como ferramenta de seleção para fungos endofíticos produtores de COVs. Dentre os 400 fungos endofíticos isolados, 11 foram capazes de crescer na presença de COVs produzidos por M. albus. Estes fungos foram identificados como Muscodor spp. (9) e Simplicillium sp. de acordo com pesquisas na base de dados UNITE usando sequências de DNA do espaçador transcrito interno (ITS). Os COVs produzidos por fungos endofíticos inibiram o crescimento dos fungos fitopatogênicos em comparação com o controle com diferentes eficácias. Os COVs produzidos por Muscodor coffeanum (COAD 1842) apresentou efeito fungicida contra A. ochraceus em grãos de café. Seis fungos endofíticos inibiram completamente o crescimento de F. verticillioides inoculado em sementes de milho. Este estudo demonstra que os fungos endofíticos produtores de compostos voláteis isolados de Coffea arabica são fontes promissoras de compostos bioativos.
Subject(s)
Aspergillus , Coffea , Fungi , FusariumABSTRACT
Ear rots caused by Fusarium spp. are among the main fungal diseases that contribute to poor quality and the contamination of maize grains with mycotoxins. This study aimed to determine the visual incidence of fungal-damaged kernels (FDKs), the incidence of two main Gibberella (a teleomorph of Fusarium) complexes (G. fujikuroi and G. zeae) associated with maize using a seed health blotter test, and the fumonisin levels, using high performance liquid chromatography, in samples of maize grains grown across 23 municipalities during the 2008/09 and 2009/10 growing seasons. Additionally, 104 strains that were representative of all of the analysed samples were identified to species using PCR assays. The mean FDK was seven per cent, and only six of the samples had levels greater than six per cent. Fusarium spp. of the G. fujikuroi complex were present in 96% of the samples, and G. zeae was present in 18% of the samples (5/27). The mean incidence of G. fujikuroi was 58%, and the incidence of G. zeae varied from 2 to 6%. FB1 was found in 58.6%, FB2 in 37.9%, and both toxins in 37.9% of the samples. The FB1 and FB2 levels were below the quantification limits for 41.3% of the samples, and the mean FB1 levels (0.66 µg/g) were higher than the mean FB2 levels (0.42 µg/g). The PCR identification separated the 104 isolates into three of the G. fujikuroi complex: F. verticillioides (76%), F. subglutinans (4%) and F. proliferatum (2%); and G. zeae (anamorph = F. graminearum) (18%). Our results confirmed the dominance of F. verticillioides, similar to other regions of Brazil, but they differed due to the relatively higher incidence of F. graminearum. Total fumonisin levels were below the maximum limit determined by current Brazilian regulations.
Subject(s)
Humans , Food Contamination , Fumonisins/analysis , Fumonisins/isolation & purification , Fusarium/growth & development , Fusarium/isolation & purification , In Vitro Techniques , Mycoses , Plant Structures , Polymerase Chain Reaction , Chromatography, High Pressure Liquid , Food Samples , Methods , Zea maysABSTRACT
The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL) and Eutroferric Red Latosol (ERL) soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05) in soils with glucose and inoculated with the fungi (except F. verticillioides), in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.
Subject(s)
Amylases/analysis , Carbohydrate Dehydrogenases/analysis , Spores, Fungal/growth & development , Mitosporic Fungi/growth & development , Enzyme Activation , Methods , MethodsABSTRACT
Este trabalho teve por objetivo isolar, identificar e selecionar isolado(s) de Fusarium spp. em sementes de pepino e verificar a sua influência na qualidade fisiológica. Foram utilizados três sublotes e os tratamentos consistiram em: testemunha absoluta; BDA + manitol - 0,8Mpa; BDA; BDA + manitol - 0,8Mpa + fungo; e BDA + fungo. A qualidade fisiológica foi avaliada através de testes em laboratório e casa de vegetação. Cinco isolados de Fusarium verticillioides foram identificados e a sua transmissão através das sementes para plantas não foi possível afirmar. A técnica da restrição hídrica foi eficiente para proporcionar infecção das sementes, sabendo que e o fungo influencia negativamente na qualidade fisiológica das sementes de pepino da cultivar Caipira.
This research had the objective of isolate, identifying and select isolates (s) of Fusarium spp. in cucumber seeds and checking the influence of this (these) isolate(s) on the physiological quality. Three batches were used and treatments utilized consisted in: absolute witness; PDA + mannitol - 0.8Mpa; PDA pure; PDA + mannitol - 0.8Mpa + fungi; and PDA + fungi. The physiological quality was assessed by tests carried out in laboratory and greenhouse. Five isolates off Fusarium verticillioides were isolated and it was not possible to affirm its transmission through seed. The hydric restriction technique was efficient to provide the infection of cucumber seeds, since this fungus affected the physiological quality of cucumber seeds of this cultivar.
ABSTRACT
In this study a total of 16 Fusarium verticillioides strains isolated from corn feed samples were characterized by fumonisin (FB) production and random amplified polymorphic DNA (RAPD). All the strains produced FB1 and FB2 with levels ranging from 2.41 to 3996.36 µg/g, and from 1.18 to 1209.91 µg/g, respectively. From the 16 F. verticillioides strains, four were identified as low (3.59 to 1289.84 µg/g), eight as intermediate (>1289.84 to 3772.44 µg/g) and four strains as high (>3772.44 µg/g) fumonisin producers. From the total of 105 loci amplified, 60 (57.14 percent) were polymorphic. RAPD analysis showed very similar patterns among low, moderate and high fumonisin-producing strains. Although RAPD markers were capable of discriminating the different F. verticillioides strains, there was no clear association between these makers and fumonisin production.
Neste estudo, 16 cepas de F. verticillioides isoladas de amostras de ração de milho foram caracterizadas com base na produção de fumonisinas (FB) e em marcadores de polimorfismos de DNA amplificado ao acaso (RAPD). Todas as cepas produziram FB1 e FB2, com níveis variando, respectivamente, de 2,41 a 3996,36 µg/g e 1,18 a 1209,91 µg/g. De acordo com a produção de fumonisinas totais (FB1 + FB2) e a distribuição por análise de quartis, do total de 16 cepas de F. verticillioides, quatro foram identificadas como baixas produtoras de fumonisinas (3,59 a 1289,84 µg/g), oito como intermediárias (>1289,84 a 3772,44 µg/g) e quatro como altas produtoras de fumonisinas (>3772,44 µg/g). Os 10 primers utilizados amplificaram 105 locos, 60 (57,14 por cento) dos quais foram polimórficos. As análises de RAPD mostraram padrões muito similares entre as cepas baixas, médias e altas produtoras de fumonisinas. Embora os marcadores RAPD tenham se mostrado capazes de discriminar as diferentes cepas de F. verticillioides, não foi detectada nenhuma associação entre estes marcadores e a produção de fumonisinas.
ABSTRACT
The study reports the occurrence of fumonisin producing Fusarium verticillioides in 90 samples of stored paddy (Oryza sativa L.)collected from different geographical regions ofKarnataka, India. Fumonisin producing F. verticillioides was identified based on micromorphological characteristics and PCR using two sets of primers. One set of primers was F. verticillioides species specific, which selectively amplified the intergenic space region of rDNA. The other set of primers was specific to fumonisin producing F. verticillioides. Eight paddy samples were positive for F. verticillioides. Eleven isolates obtained from these samples were capable of producing fumonisin.
O estudo relata a ocorrência de Fusarium verticillioides produtor de fumonisina em 90 amostras de arroz armazenado, coletado de diferentes regiões geográficas de Karnataka, Índia. F. verticillioides produtor de fumonisina foi identificado baseado em características micromorfológicas e PCR empregando dois sets de primers. Um dos sets era F. verticillioides especie-específico, que amplificava seletivamente a região do espaço intergênico do rDNA. O outro set de primers era especifico para F. verticillioides produtor de fumonisina. Oito amostras de arroz foram positivas para F. verticillioides. Onze isolados obtidos dessas amostras foram produtores de fumonisina.
Subject(s)
Fumonisins/analysis , Fusarium/genetics , Fusarium/isolation & purification , In Vitro Techniques , Mycotoxins/analysis , Mycotoxins/isolation & purification , Oryza/genetics , Polymerase Chain Reaction , Food Samples , Methods , MethodsABSTRACT
Onychomycosis is caused by dermatophytes usually, but some species of non-dermatophytic molds and yeasts are also associated with invasion of nails. Fusarium (F.) verticillioides is a non-dermatophytic mold, which are common soil saprophytes. Onychomycosis due to Fusarium species almost always involves great toe nails and occurs infection easily in the presence of trauma, onychodystrophy and prior onychomycosis. We report a first case of onychomycosis caused by Fusarium verticillioides in a 48-year-old man. He presented with proximal white superficial onychomycosis for 3 months. Fungal culture showed whitish cottony colonies and microscopic examination of colonies showed microconidia in a chain from monophialide. We diagnosed onychomycosis caused by Fusarium verticillioides and treated by itraconazole 200 mg daily and 5% amorolfine nail lacquer for 3 months.
Subject(s)
Humans , Middle Aged , Arthrodermataceae , White People , Fungi , Fusarium , Itraconazole , Lacquer , Morpholines , Nails , Onychomycosis , Soil , Toes , YeastsABSTRACT
Fusarium species, which are common soil saprophytes, have been implicated as a pathogen causing a variety of opportunistic infections such as keratitis, onychomycosis, burn wound infection or disseminated systemic infections in immunocompromised hosts. We report a case of cutaneous infection caused by F. verticillioides in a 59-year-old man, with a long history of diabetes mellitus and alcoholic liver cirrhosis. He presented with a few papules and fluctuant cutaneous nodules on both forearms traumatized by woodstick 4 months ago. The diagnosis of F. verticillioides infection was established by histopathologic examination and finding of tissue cultures.